Immunofluorescence was performed following standard procedures24 (link). The primary antibody was anti-phospho-histone-3 (PH3, 1:2000; Ab14955, Abcam), and the secondary antibody was anti-mouse Alexa Fluor 555 IgG (Invitrogen). Nuclear DNA was stained with Hoechst 33342 (Sigma). Pictures were taken with an Eclipse 80i microscope and processed with the NIS Elements Advances Research software (Nikon).
Nis elements advances research software
Manufactured by Nikon
Sourced in Japan
NIS Elements Advances Research software is a comprehensive imaging and analysis solution for life science research. It provides a range of tools and functions to capture, process, analyze, and manage digital microscopy data. The software supports a variety of imaging techniques and file formats, allowing researchers to work with diverse imaging data.
Automatically generated - may contain errors
Lab products found in correlation
Ab14955, by Abcam (4 mentions)
Eclipse 80i microscope, by Nikon (4 mentions)
Hoechst 33342, by Merck Group (4 mentions)
Alexa fluor 555 igg, by Thermo Fisher Scientific (2 mentions)
Af835, by R&D Systems (2 mentions)
Anti mouse igg alexa fluor 555, by Thermo Fisher Scientific (1 mentions)
A31572, by Thermo Fisher Scientific (1 mentions)
A21422, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 igg, by Thermo Fisher Scientific (1 mentions)
4 protocols using nis elements advances research software
1
Immunofluorescence Staining for PH3
2
Immunofluorescence Staining for p-H3 and Caspase-3
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Immunofluorescence was performed as described in previous studies24 (link). Cells were incubated with phospho-histone H3 (p-H3) (ab14955, Abcam) and caspase-3 (AF835, R&D Systems) antibodies. Secondary antibodies anti-mouse and anti-rabbit were Alexa Fluor 555 IgG (A21422, A31572, respectively; Invitrogen). Nuclear DNA was stained with Hoechst 33342 (Sigma). Pictures were taken with an Eclipse 80i microscope and processed with the NIS Elements Advances Research software (Nikon).
3
Immunofluorescent Imaging of Apoptosis Markers
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It was performed as described in previous studies (22 (link)). Cells were incubated overnight at 4°C with primary antibody for p-H3 (ab14955 Abcam, 1/2000) and active caspase-3 (AF835 R&D Systems, 1/500). Then, cells were incubated for 1 hour with secondary Alexa Fluor 555 IgG anti-mouse (A21422 Invitrogen, 1/500) or anti-rabbit (A31572 Invitrogen, 1/500) antibodies at room temperature in darkness. For nuclear DNA staining, Hoechst 33342 (Sigma) was used. Pictures were taken with an Eclipse 80i microscope and processed with the NIS Elements Advances Research software (Nikon).
4
Immunofluorescence for SOX2 and pH3
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Immunofluorescence was performed as described in previous studies [26 (link)]. Cells were incubated with SOX2 (AB5603 Millipore, Burlington, MA, USA) and phospho-histone3 (pH3) (ab14955 Abcam, Cambridge, UK) antibodies. Secondary antibodies of anti-mouse Alexa Fluor 555 IgG (Invitrogen, Carlsbad, CA, USA) and anti-rabbit Alexa Fluor 488 IgG (Invitrogen, Carlsbad, CA, USA) were used. Nuclear DNA was stained with Hoechst 33342 (Sigma, St. Louis, MO, USA). Pictures were taken with an Eclipse 80i microscope and processed with the NIS Elements Advances Research software (Nikon, Tokyo, Japan).
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